DURING HYPOXIC CEREBRAL ISCHEMIA, ghCOse Cannot MATERIALS AND METHODS Tissue Dissociation and Culture Forebrain cultures were prepared from El6 rat embryos as previously described(8). Cells 2 x lo6 were plated on poly-Llysine-coated 25-mm round cover slips placed in 35-mm Corning dishes.The cultures were kept at 37Â°C in 5% COPhumidified air. Cultures were exposedto after 8-10 days in vitro (DIV). Measurement of Intracellular pH be oxidized through the Krebs cycle, resulting in the local production and accumulation (pK, 3.84). After 30 min of incomplete ischemia, brain lactate concentration rises from a normal of 1-16 pmol/g in fasted animals and may reach levels of 34 pmol/g in hyperglycemic rats (25). Concomitantly, the brain interstitial (external) pH (pH,) falls from 7.3 to 6.8 in fasted animals and to as low as 5.8 in hyperglycemic animals (9, 17). These degrees osis are sufficient to induce cellular damage in vitro (7, 8, 16, 18). On this basis, it has been proposed that lactic production may mediate the conversion of selective neuronal damage into tissue infarction (20). We have previously demonstrated that -induced cell injury in vitro occurs as a function of both the intensity and duration of intracellular ification. An intracellular pH (pHi)
AJP - Regulatory, Integrative and Comparative Physiology – The American Physiological Society
Published: Aug 1, 1993
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